MS2 tagging - Wikipedia
MS2 tagging is a technique based upon the natural interaction of the MS2 bacteriophage coat protein with a stem-loop structure from the phage genome, [1] which is used for biochemical purification of RNA-protein complexes and partnered to GFP for detection of …
The RNA binding site of bacteriophage MS2 coat protein
The coat protein of the RNA bacteriophage MS2 binds a specific stem-loop structure in viral RNA to accomplish encapsidation of the genome and translational repression of replicase synthesis. In order to identify the structural components of coat protein required for its …
Methods to study RNA–protein interactions - Nature
2019年2月25日 · Both MS2 hairpin RNA and MS2 coat protein are expressed in the same cell and form a stable complex, enabling the fusion MS2 coat protein to be used as a handle to purify the...
An improved imaging system that corrects MS2-induced RNA
2022年11月10日 · The MS2 and MS2-coat protein (MS2-MCP) system is widely used to study mRNA localization in living cells 2. In this approach, an array of MS2 binding sites (MBS) is inserted downstream...
An improved MS2 system for accurate reporting of the mRNA life cycle
The MS2-MCP system allows imaging multiple steps of the mRNA life cycle with high temporal and spatial resolution. However for short-lived mRNAs, the tight binding of the MS2 coat protein (MCP) to the MS2 binding sites (MBS) protects the RNA from ...
New Generations of MS2 Variants and MCP Fusions to Detect …
To visualize single mRNAs in living cells, 24 MS2 stem-loops are used to tag an RNA of interest, which is then detected by co-expression of a specific RNA-binding protein, the MS2 coat protein (MCP), fused to fluorescent proteins (FP, i.e., eGFP, mCherry, tdTomato, HALO, photoactivatable proteins) [1, 39].
An improved MS2 system for accurate reporting of the mRNA …
2017年11月13日 · Here, we describe a reporter system (MBSV6) with reduced affinity for the MCP, which allows mRNA degradation while preserving single-molecule detection determined by single-molecule FISH (smFISH)...
Crystal structure of the MS2 coat protein dimer: implications for …
The coat protein from the MS2 bacteriophage is a member of a group of small proteins that bind RNA in a multifunctional manner in related RNA bacteriophages. These spherical viruses infect Escherichia coli, and their genomic RNA serves as mRNA for synthesis of viral proteins.
Identification of mRNA-interacting factors by MS2-TRAP (MS2 …
Here, we provide a detailed protocol to tag mRNAs with MS2 hairpins and then affinity-purify trans-binding factors (RBPs, ncRNAs) associated with the MS2-tagged mRNA. This method, termed MS2-TRAP, permits the systematic characterization of ribonucleoprotein (RNP) complexes formed on a given mRNA of interest.
Complementation of RNA binding site mutations in MS2 coat protein …
1996年6月15日 · The coat protein of bacteriophage MS2 functions as a symmetric dimer to bind an asymmetric RNA hairpin. This implies the existence of two equivalent RNA binding sites related to one another by a 2-fold symmetry axis. In this view the symmetric binding site defined by mutations conferring the repress …